The regulatory region and the signal peptide sequence of the sacB gene has been amplified by PCR using Bacillus subtilis chromosomal DNA as template, and an inducible secretion vector has been developed based on this sequence, which was ligated with Bacillus subtilis alkaline proteinase gene. Transform Bacillus subtilis DB403 with this vector, and the expression of the inserted Bacillus subtilis alkaline proteinase gene can be induced by addition of sucrose into the medium.

Abstract: Objective To explore the influencing factors of serum HBeAg loss in patients with chronic hepatitis B (CHB) and and provide evidence for effective treatment of CHB. Methods A follow-up cohort of HBeAg-positive CHB patients was established in the the Infectious Diseases Outpatient Clinic of hospital. Regular follow-up and laboratory test indicators were collected to analyze the changes of serum HBeAg in HBeAg-positive CHB patients during the follow-up period. The subjects were divided into the case group (serum HBeAg loss) and the control group (serum HBeAg not loss) according to whether serum HBeAg loss occurred. The baseline data characteristics of the two groups were analyzed and compared, and the influencing factors of serum HBeAg loss were analyzed by Cox univariate and multivariate regression. Results A total of 634 HBeAg-positive CHB patients were enrolled, with a total follow-up of 2 570.01 person-years. Among them, 237 cases of serum HBeAg loss occurred, with the mean follow-up time of 40.92 months, and the rate of HBeAg loss was 9.22/100 person-years. There were significant differences in HBV family history, antiviral therapy, baseline WBC, PLT, ALT, AST, T˗Bil, GGT, AFP, quantitative HBsAg and quantitative HBeAg between serum HBeAg loss group and serum HBeAg not loss group (P<0.05). Cox regression analysis showed that family history of HBV (HR 0.68, 95％CI:0.50-0.92, P=0.012), ALT (HR2.06, 95％CI:1.52-2.79, P<0.001), quantitative HBsAg (HR 0.68, 95％CI:0.48-0.95, P=0.024), quantitative HBeAg (HR 0.48, 95％CI:0.31-0.74, P=0.001) were independent influencing factors for HBeAg loss in HBeAg-positive CHB patients. Conclusions HBeAg-positive CHB patients without family history of HBV, initial ALT≥80 U/L, quantitative HBsAg<1 000 IU/ml, quantitative HBeAg<1 000 C.O.I are more likely to have serum HBeAg loss.

Objective To investigate the effect of Ulinastatin on the delivery of cytokines in patients with septic shock.Methods It was a prospective and controlled clinical study.Seventy-eight patients with septic shock were randomly divided into control group and treatment group and thirty-nine in every group.Patients in treatment group received Ulinastatin 200 000 units intravenous everyday for 3 days,while those in control group received equal volume of normal saline as placebo.At different time points (at 24 th,48 th,72 th hour after start of treatment),the levels of tumor necrosis factor-alpha (TNF-?),interleukin-1 (IL-1),interleukin-6 (IL-6 ),interleukin-8 (IL-8) and superoxide dismutase (SOD) in serum were assayed.Results In comparison with control group,the levels of TNF-?,IL-1,IL-6,IL- 8 of treatment group decreased markedly (P＜0.05,P＜0.01) at different time points,whereas the level of SOD was higher markedly (P＜0.05,P＜0.01) at various time points.Conclusion Ulinastatin has protective effect on patients with septic shock through decreasing the levels of TNF-?,IL-1,IL-6,IL-8 and increasing in the level of SOD.

This study was purpose to investigate the effects of CD147 on the invasiveness of leukemia cells U937. The experiments were divided into 4 groups: control group, LPS group, CD147mAb group and LPS+CD147 mAb group. Cells were treated by lipopolysaccharide (LPS) or anti-CD147 monoclonal antibody, and the expression of CD147 and MMP-2, -9, the invasive potential of the cells in vitro and ex vivo, as well as the invasion of the implanted tumors in SCID mice were analysed by RT-PCR, FCM, gel zymography and invasion test in vitro respectively. The results showed that the expression of CD147 was elevated by the induction of LPS, and the enhanced expression of CD147 on U937 cells increased the production and secretion of MMP-2 and MMP-9 as measured by reverse transcription-PCR and gel zymography. An increased number of LPS-induced cells invading through a reconstituted basement membrane were observed by invasion assays. These responses were down-regulated after blocking CD147 with anti-CD147 antibody. At 30 days after intravenous injection of LPS pretreated U937 cells to SCID mice human U937 cells were found in the bone marrow and lung of the mice, indicating the invasion of the tumor cells. And overexpressions of CD147, MMP-2 and MMP-9 were found in the lung tissue of the mice injected with LPS-treated but not anti-CD147 antibody treated tumor cells. It is concluded that overexpression of CD147 on U937 cells may increase the secretion and activation of MMP-2 and MMP-9 and thus promote the invasiveness of the tumor cells.
Animals ; Antibodies, Monoclonal ; pharmacology ; Basigin ; genetics ; immunology ; metabolism ; Female ; Humans ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Mice ; Mice, Nude ; Mice, SCID ; Neoplasm Invasiveness ; RNA, Messenger ; genetics ; metabolism ; U937 Cells

Accidents, Occupational ; statistics & numerical data ; China ; epidemiology ; Humans

OBJECTIVESTo explore the clinical application of anti-human seminal plasma phospholipase A2 (PLA2) monoclonal antibody (McAb) for male infertility.

METHODSEnzyme-linked immunoabsorbent assay (ELISA), immunocytochemistry(ICC), as well as flow cytometry (FCM) analysis were established using two strains anti-human seminal plasma PLA2 McAb prepared by our laboratory to detect the PLA2 content in human seminal plasma and the anterior head region of spermatozoa, respectively. Then the PLA2 content in male infertile patients were compared with that in normal control with fertility. The seminal routine analysis was performed by computer-assisted semen analysis (CASA).

RESULTSThe PLA2 content of infertile groups were (31.13 +/- 14.49) ng/ml in azoospermic patients, (17.71 +/- 12.45) ng/ml in oligospermic patients and (16.46 +/- 11.31) ng/ml in patients with normal sperm density, which were all higher than that of normal controls [(8.09 +/- 3.15) ng/ml, P < 0.01]; There was significantly negative correlation between PLA2 content in seminal plasma and sperm density(r = -0.602, P < 0.05), while there was insignificant correlation between PLA2 and sperm motility or percentage of motility. The PLA2 content in the anterior head region of spermatozoon of male infertile groups was significantly lower than that of normal controls by ICC and FCM(P < 0.01).

CONCLUSIONSPLA2 in human seminal plasma is closely related to male fertility, and the PLA2 deficiency in the head of spermatozoa may be one of the reasons causing male infertility. The methods detecting PLA2 content in seminal plasma and the head of spermatozoa can provide powerful evidences for exploring the mechanism of male infertility.

Adult ; Humans ; Infertility, Male ; enzymology ; Male ; Phospholipases A ; analysis ; Phospholipases A2 ; Semen ; enzymology

OBJECTIVETo investigate the changes of the goblet cells in the intestine during the restitution process of the gut barrier after hemorrhagic shock.

METHODSForty-nine Sprague-Dawley rats with body weight of 250-300 g were divided into control group (n=7) and experimental group (n=42). Rats in the experimental group was further divided into 6 groups (n=7 each) according to different time point at 1, 3, 6, 12, 24, and 36 hours after hemorrhagic shock resuscitation. The specimens from ileum tissue were taken to observe the morphological chan ges of the intestinal mucosa. The number of goblet cells was determined by light microscope and/or electron microscope. The contents of trefoil factor family 3 (TFF3) of goblet cells were examined using GC-9A gas chromatographic instrument.

RESULTSAfter hemorrhagic shock, mucosal epithelial injury was obvious in the small intestine. Tissue restitution was found after 3 hours, and mostly established after 12 hours. Following tissue restitution,the denuded mucosal surface was covered intensively by goblet cells. The number of goblet cells on the intestinal mucosa was reduced significantly from 243+/- 13 at 1 h to 157+/- 9 at 24 h (r=- 0.910, P< 0.01), and returned to normal level at 36 h. In the experimental group, the content of TFF3 in the intestinal mucosa increased significantly at 12 hours, decreased, but was still higher at 24 hours (t=3.24, P< 0.05).

CONCLUSIONSThe goblet cells play a key role in the restitution of intestinal mucosa. High expression of TFF3 may facilitate the intestinal mucosal restitution in the early phase.

Animals ; Goblet Cells ; metabolism ; Ileum ; cytology ; Intestinal Mucosa ; cytology ; metabolism ; pathology ; Neuropeptides ; metabolism ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; metabolism ; Trefoil Factor-3

Objective Constructing physiological-psycho-social three dimensional aging index system, and scale to provide a basis for evaluating the effects of human aging and anti-aging measures. Methods Evaluation, modification and determination of the second and third level indexed by Delphi and principal component analysis and other methods. The weight of each index was weighted by the analytic hierarchy process, and the reliability and validity were calculated on the basis of the preliminary scale. A large sample of empirical studied to determine the aging of different age groups standardized score distribution table. The best model for judging functional age was sought, based on a variety of biostatistical models. Results The aging scale was constructed by three levels, the first level included 3 indicators, the second level had 10 indicators, the third level included 51 indicators, and adopted by expert certification the national society. 3 184 valid questionnaires were obtained through epidemiological investigation, the average total score of aging was (46.93±11.07) points, and the scores of aging were positively correlated with age(r=0.785, P<0.001). The average scores of aging in each dimension were different and the difference was statistically significant (all P<0.001), and each dimension had significant difference with age. The growth model (R2=0.635) was selected, the curve of the relationship between the score of aging and age was fitted. Conclusions The three-dimensional human aging scale was scientific, feasible, and has good reliability and validity. The empirical results show that the comprehensive score of aging increase with age, and show certain characteristics and laws on the curve.

OBJECTIVETo study the therapeutic effect of bleomycin-A5 injection under eletrolaryngoscope for treatment of large laryngopharyngeal and laryngeal hemangioma.

METHODSIntratumor bleomycin-A5 injection under eletrolaryngoscope was performed in 18 cases of large laryngopharyngeal and laryngeal hemangioma with surface anaesthesia, for totally 7 to 14 (mean 10.2) injections in each case.

RESULTSTwelve patients were cured and 6 showed obvious improvement. Follow-up of the patients for over one year found no recurrence of the hemangioma.

CONCLUSIONBleomycin-A5 injection is a minimal invasive procedure for treatment of large laryngopharyngeal and laryngeal hemangioma, causing less pain and better preserving the laryngeal function without the necessity of tracheotomy.

Adolescent ; Adult ; Aged ; Antibiotics, Antineoplastic ; administration & dosage ; Bleomycin ; administration & dosage ; analogs & derivatives ; Female ; Follow-Up Studies ; Hemangioma ; drug therapy ; Humans ; Injections, Intralesional ; methods ; Laryngeal Neoplasms ; drug therapy ; Laryngoscopes ; Male ; Middle Aged ; Pharyngeal Neoplasms ; drug therapy